Project Summary/Abstract: Next Generation Immunoanalysis Core Research on immune reactions in human skin and human inflammatory diseases has been hampered by the relative paucity of reagents, such as tetramers, that are available for human-based research. However, there has been a revolution in the last five years in the tools available for human immune profiling. These techniques have opened the door to comprehensive study of immune reactions in human skin and blood. The purpose of this Core is to make two critical groundbreaking techniques, cytometry by time of flight (CyTOF) and next- generation high throughput T cell receptor sequencing (HTS), available to skin disease researchers at any institution with the goal of accelerating translational research in human inflammatory skin diseases. CyTOF combines immunostaining with mass spectroscopy and allows measurement of up to 45 different markers in single sample, with no signal overlap and no need for compensation. CyTOF instruments are located in a small number of major academic medical centers. We provide access to the Longwood Medical Area CyTOF facility and also provide services that will make CyTOF accessible, including staining protocols, validated panels of CyTOF antibodies, custom conjugated CyTOF antibodies, advice on experimental design and data analysis, and all services needed for outside investigators to send their cells to the Center for analysis. HTS is a powerful technique that allows one step comprehensive profiling of T cells in tissues and the ability to identify and track pathogenic T cell clones across various tissues, including blood and skin. We provide comprehensive support in experimental design, DNA preparation protocols, DNA sample preparation services, shipping, and data analysis assistance that will enable human skin disease researchers to rapidly use this technique to profile immune skin immune responses. We also offer custom staining for pathogenic T cell clones using a combination of HTS analyses and immunostaining of tissue sections with commercially available TCR V? antibodies. These combined techniques allow selective identification and study of particular T cell clones. The research community potentially includes any individual wishing to carry out human skin disease research; we have included 24 projects from investigators who wish to utilize Center services. 18 users would like to utilize the Immunoanalysis Core for their projects. 11 are from outside institutions, six have never worked in human skin disease before and six have worked primarily in mouse models previously, with little or no prior work in humans. Eight of these projects are described in detail in this component and other projects are described fully in the two other Resource Core portions of the application. In summary, the Next Generation Immunoanalysis Core will make CyTOF and HTS, two cutting edge analytic techniques, available to researchers at any institution who wish to carry out human tissue based skin disease research.